CN111172062B - Sphingobacterium multivorum and application thereof - Google Patents
Sphingobacterium multivorum and application thereof Download PDFInfo
- Publication number
- CN111172062B CN111172062B CN201911378511.6A CN201911378511A CN111172062B CN 111172062 B CN111172062 B CN 111172062B CN 201911378511 A CN201911378511 A CN 201911378511A CN 111172062 B CN111172062 B CN 111172062B
- Authority
- CN
- China
- Prior art keywords
- straw
- cellulose
- sphingosine
- multivorum
- sphingobacterium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The embodiment of the invention discloses application of Sphingobacterium multivorum cellulose decomposition bacteria in cellulose degradation. The embodiment of the invention also discloses a cellulose degrading enzyme preparation which contains the fermentation liquor of the sphingosine bacillus. The sphingosine bacillus multivorus provided by the embodiment of the invention has a high-efficiency degradation function on crop straws in soil and water, so that the crop straws in the soil and water can be rapidly and efficiently degraded.
Description
Technical Field
The embodiment of the invention relates to the technical field of microorganisms, and in particular relates to sphingobacterium multivorum and application thereof.
Background
The safety problem of agricultural products and food is the first problem of concern to the national civilization, and with the consumption of resources, people are confronted with the problems of land resource shortage, excessive use of chemical fertilizers and pesticides, serious pollution of heavy metals and organic matters, improper disposal of straw wastes and the like, great potential safety hazards are caused to agricultural products and food, and more serious environmental pollution is possibly caused. The crop straw is used as a natural associated product of agricultural products, and after being decomposed, a large amount of biological organic matters and nutrient elements such as nitrogen, phosphorus, potassium and the like are released. Straw in-situ returning is one of important means for comprehensive utilization of straw, and if the level of returning to the field in developed countries can be reached, namely 75% -80% returning to the field is achieved, not only is 3500 million tons of chemical fertilizer saved every year, but also the problem of air pollution caused by straw burning can be avoided. The in-situ straw returning process includes crushing the straw in situ while harvesting, returning the straw to soil directly via ploughing, and converting the organic matter in the straw into organic matter for storing in soil after the action of soil microbes. The direct returning of the straws to the field can increase the organic matter content of the soil, improve the volume weight and permeability of the soil and increase the water content of the soil, and is an important means for economic and efficient and promoting the sustainable development of agriculture.
At present, fungi are generally adopted to degrade cellulose in straw treatment, but in the actual fermentation treatment process, the bacteria have larger comparative area and can quickly absorb soluble substrates into cells, and the quantity of the soluble substrates is more than that of actinomycetes and fungi. In addition, the microbial inoculum used for the existing cellulose decomposition is the mixture of various microbial inocula, the effect is poor, and the treated straw is single.
Disclosure of Invention
Therefore, the embodiment of the invention provides a sphingosine bacillus polyphylla and application thereof, and aims to solve the problem that no strain capable of effectively degrading cellulose exists in the prior art.
In order to achieve the above object, the embodiments of the present invention provide the following technical solutions:
sphingobacterium multivorum cellulolytic bacteria, which was deposited in the general microbiological center of the China Committee for culture Collection of microorganisms at 5.22.2015, 5.22.D., with the deposit number of CGMCC No.10851, No. 3 of Ministry of microbiology of China, Ministry of sciences, Ministry of China, and the deposit number of CGMCC No. 10851.
The invention also provides a cellulose degrading enzyme preparation which contains the fermentation liquor of the sphingosine bacillus.
Preferably, the cellulose degrading enzyme preparation comprises cellulase and/or hemicellulase and/or laccase and/or manganese catalase and/or lignin catalase.
Preferably, the cellulose-degrading enzyme preparation has a degradation substrate comprising sodium carboxymethyl cellulose and/or lignin.
The application of Sphingobacterium multivorum decomposing bacteria in degrading cellulose also belongs to the protection scope of the embodiment of the invention.
Preferably, the cellulose is straw.
Preferably, the straw is at least one of rice straw, wheat straw and corn straw.
In the embodiment of the invention, the degradation of the straws in the environment can be carried out under the condition that the sphingosine bacillus multivorus can survive. Wherein, the term "sphingosine dormitogenes are viable" means that at least more than 5%, preferably at least more than 10%, more preferably at least more than 60% of the biomass is viable in an environment containing straw.
In the present embodiment, the term "conditions under which sphingosine bacteria can survive" refers to conditions that at least include the most basic conditions under which sphingosine bacteria can survive, such as temperature, nutrient sources, etc., and are well known to those skilled in the art, and thus, are not described herein again.
In order to further improve the viability of said sphingosine bacilli in the environment, it is also possible to add to the environment in need of remediation a source of nutrients suitable for the growth of said sphingosine bacilli, such as glucose and the like.
The straw-containing environment may include any straw-containing environment, for example, the straw-containing environment may include soil or a body of water containing straw.
According to the embodiment of the invention, the form of the sphingosine pluriovorans added to the environment containing straws is not particularly limited as long as the sphingosine pluriovorans can be ensured to act in the environment containing straws and effectively degrade the straws after the addition. The sphingosine multivorum is added, for example, in the form of a bacterial liquid, a pellet of the bacterial, or a dry powder.
The sphingosine bacillus multivorus provided by the invention has dosage and time effects on the degradation of straws, for example, the degradation rate of the straws can be increased along with the increase of the concentration of the sphingosine bacillus multivorus or the prolonging of the time in a certain range.
The number of added sphingosine bacillus multivorus is not particularly limited in the embodiment of the invention, and can be determined according to the content of straw in the environment containing straw, for example, when the content of straw in the environment is higher or the environment is less favorable for the survival of the sphingosine bacillus multivorus, the inoculation amount of the sphingosine bacillus multivorus can be increased; when the straw content in the environment is low or the influence of the environment on the survival of the sphingosine multivora is small, the inoculation amount of the sphingosine multivora can be reduced.
According to the embodiment of the invention, when the environment containing the straws is soil, in order to further promote the degradation efficiency of the sphingosine multivorum provided by the invention on the straws, the water content in the soil is preferably controlled to be at least 15 wt%, and preferably 20-30 wt%.
In the present invention, the viable cells of sphingosine bacillus can be further separated from the culture medium, and the method for separating is not particularly limited as long as the cells can be enriched from the culture medium, and for example, the separation can be achieved by centrifugation and/or filtration, and the conditions for centrifugation and filtration can be known conditions, and the present invention is not described herein again.
In the embodiment of the invention, the cellulase preparation can be in the form of a bacterial liquid or a dry powder, and can be selected by a person skilled in the art according to actual needs. The bacterial liquid may be, for example, a bacterial liquid of Sphingobacterium multivorum obtained by culturing the above-mentioned bacterial liquid medium. The dry powder may be, for example, a dry powder obtained by freeze-drying the bacterial liquid of sphingosine bacillus multivorus cultured up to the logarithmic phase.
The embodiment of the invention has the following advantages:
the sphingosine multivorum (Sphingobacterium multivorum) of the embodiment of the invention has the activities of producing cellulase and hemicellulase, and producing manganese peroxidase, laccase and lignin catalase; the sphingosine multivorum has a high-efficiency degradation function on crop straws in soil and water, can quickly decompose the straws, release nutrients, provide nutrients for other microorganisms in the environment, degrade the straws together with the sphingosine multivorum, improve the soil microecology, further improve the degradation rate of the straws, and have high economic benefit and social benefit.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
FIG. 1 is a graph showing the streaked culture results of a solid culture medium of Sphingobacterium multivorum according to an embodiment of the present invention;
FIG. 2 is a diagram showing the result of comparison between the sequence of the 16S rDNA gene of Sphingobacterium multivorum provided in the embodiment of the present invention and the sequence of the 16S rDNA gene of the existing strain in GenBank.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the present example, the amount of enzyme required to produce 1mol of glucose from a substrate per minute is defined as one enzyme activity unit (U).
In the embodiment of the invention, the formula of the bacterial liquid culture medium is as follows: 10g/L of peptone, 5g/L of yeast powder and 10g/L of sodium chloride, and the pH value is 6.8-7.0.
The formula of the culture medium for producing the cellulase by the straws is as follows: 10-15g/L straw powder, 0.8-1.5g/L potassium dihydrogen phosphate, 0.3-0.5g/L dipotassium hydrogen phosphate, 0.3-0.5g/L magnesium sulfate, 0.010-0.015g/L calcium chloride and 0.3-0.5g/L ammonium nitrate.
The formula of the culture medium for producing ligninase by straws is as follows: 0.2-0.3g/L ammonium tartrate, 0.2-0.3g/L dipotassium hydrogen phosphate, 0.03-0.06g/L magnesium sulfate and 0.01-0.02g/L calcium chloride.
The inorganic salt liquid culture medium comprises the following components in percentage by weight: 0.8-1.2g/L ammonium sulfate, 0.3-0.8g/L dipotassium hydrogen phosphate, 0.3-0.8g/L monopotassium phosphate, 0.2-0.5g/L magnesium sulfate and 20-25g/L agar powder, and the pH value is 6.0-7.5.
The LB solid culture medium formula is: 10g/L of peptone, 5g/L of yeast powder, 10g/L of sodium chloride and 20-25g/L of agar powder, wherein the pH value is 6.8-7.0.
The formula of the inorganic salt screening solid culture medium containing straw powder is as follows: 4.0-5.0g/L straw powder, 0.8-1.2g/L ammonium sulfate, 0.3-0.8g/L dipotassium hydrogen phosphate, 0.3-0.8g/L potassium dihydrogen phosphate, 0.2-0.5g/L magnesium sulfate and 20-25g/L agar powder, and the pH value is 6.0-7.5.
Example 1 isolation and identification of cellulolytic bacteria
First, separation of cellulolytic bacteria
The Sphingobacterium multivorum of the embodiment of the present invention was isolated from the soil of Xuancheng, Anhui. The method for isolating Sphingobacterium multivorum adopts a method for screening new strains, which is conventional in the field, and combines soil circulation separation with dilution plate purification. The method comprises the following specific steps:
200g of soil collected from Xuan city of Anhui and 10g of straws are weighed and placed into a circulation enrichment device. Pumping 1500mL of 121 deg.C sterilized inorganic salt liquid culture medium for 15min into fluidized bed with peristaltic pump at 170rpm with air pump displacement of 2 L.min-1Supplying inorganic salt culture medium to the fluidized bed at intervalsAnd adding the circulating fluid periodically according to the evaporation condition of the circulating fluid in the enrichment process of the solution. Taking 100 μ L of the circulation fluid every 2d, and diluting the samples 10 times respectively2、104、106、108Respectively coating 100 mu L of the strain on an LB solid culture medium, placing the LB solid culture medium in a thermostat at 30 ℃ for culturing for 48 hours, streaking and separating to obtain a single bacterial colony, and inoculating the obtained single bacterial strain on an inorganic salt screening solid culture medium containing straw powder for secondary screening. A cellulose decomposing bacterium which can decompose cellulose and has high-efficiency degradation capability on straws is obtained from the purified strains.
Identification of cellulolytic bacteria
1. Morphological characteristics of bacterial colony
The cellulose decomposition bacteria obtained by the previous separation step are subjected to streak culture, and as shown in figure 1, the cellulose decomposition bacteria colonies are turbid, semitransparent, glossy, round and convex, the surfaces of the cellulose decomposition bacteria colonies are smooth and moist, and the edges of the cellulose decomposition bacteria colonies are tidy.
2. 16S rDNA sequence analysis
The bacterial seed solution used by the cellulolytic bacteria of the embodiment of the invention is prepared by sterilizing the bacterial liquid culture medium at 121 ℃ for 15min and cooling to room temperature. Inoculating 3 vol% of cellulose decomposition bacteria liquid into 200mL of the sterilized bacteria liquid culture medium, and culturing for 48h under the culture conditions of 150-240rpm and 25-37 ℃ to obtain the cellulose decomposition bacteria seed liquid.
Extraction of DNA from cellulolytic bacteria A genomic DNA extraction kit (TIANGEN, DP302) from the strain was used and the genomic DNA of the strain was extracted according to the instructions in the kit.
The 16S rDNA gene of cellulolytic bacteria was PCR amplified using universal primers 27F and 1492R as shown in Table 1. The PCR reaction system is 50 μ L: 2 × Taq PCR mix 25 μ L, 27F 1.0 μ L, 1492R 1.0 μ L, ddH2O 23.0μL。
The PCR reaction program is: denaturation at 94 deg.C for 5min, denaturation at 94 deg.C for 30S, denaturation at 57 deg.C for 30S, denaturation at 72 deg.C for 1min for 30S, and extension at 72 deg.C for 10min, and preserving at 4 deg.C.
The PCR amplification products were detected using 0.8% agarose gel electrophoresis, spotted: 3 μ L of PCR product +1 μ L of LGel-Red, electrophoresis conditions were 150V 15 min. And (5) sending the sample with positive detection result to the Huada gene sequencing.
TABLE 1
The obtained 16S rDNA gene of cellulolytic bacteria has a full-length sequence of 1421bps, a nucleotide sequence of SEQ ID NO 3, and the comparison result of GenBank is analyzed, so that the 16S rDNA gene sequence of the strain has 99 percent of homology with a part of the sequence of Sphingobacterium sp.16S rDNA, and the bacterium can be determined to be Sphingobacterium sp. The strain has 99% homology with 16S rDNA sequence of Sphingobacterium Multivorum strain in matching with known strains. The alignment result of the 16S rDNA gene full-length sequence (1421bps) of the strain in GenBank is shown in FIG. 2. The cellulolytic bacteria is identified as Sphingobacterium multivorum, and is deposited in the general microorganism center of China Committee for culture Collection of microorganisms, 5-month and 22-month in 2015, with the deposition address: the collection number of the microorganism research institute of Chinese academy of sciences, No. 3 Xilu No.1 of Beijing, Chaoyang, and the institute of microorganisms is CGMCC No. 10851.
Example 2 cellulase and hemicellulase Activity produced by Sphingobacterium multivorum
1. Preparation of crude enzyme solution
Inoculating sphingosine bacillus polyphaga to a straw cellulase production culture medium for culture. After culturing for 6 days, 7mL of liquid is taken out and put in a centrifuge tube, and centrifuged for 10min at 4 ℃ and 5000rpm, and the supernatant is taken as crude enzyme liquid.
2. Determination of cellulase Activity
Taking 4 clean and dried test tubes, numbering 1-4, adding 1.5mL of 0.5% sodium carboxymethylcellulose solution into each test tube, adding 2mL of DNS solution into the test tube No.1, and preheating the 4 test tubes in a constant-temperature water bath at 50 ℃ for 10 min; 0.5mL of the above crude enzyme solution was added to each tube, and the reaction was carried out for 30min while maintaining a constant temperature water bath at 50 ℃. Immediately after the reaction was terminated, a 2ml of a dns solution was added to the other tube to terminate the reaction. And (3) placing 4 test tubes in a boiling water bath for 10min, cooling to normal temperature, diluting to 20mL with deionized water, and fully and uniformly mixing. And (4) zero setting by taking the test tube 1 as a blank, measuring the absorbance of the liquid in other test tubes at the wavelength of 540nm, and calculating the corresponding cellulase activity.
The method for determining the cellulase activity by DNS takes the test tube 1 as a blank to be adjusted to zero, the absorbance of liquid in other test tubes under the wavelength of 540nm is determined, and the corresponding hemicellulase activity is calculated.
The results show that the endo-cellulose enzyme activity and the hemicellulase activity of the sphingosine bacillus multivorus are respectively 0.35 U.mL-1And 3.56 U.mL-1. Therefore, the sphingosine multivorum of the embodiment of the invention can efficiently degrade cellulose.
Example 3 manganese peroxidase, laccase and Lignin Catalase production Activity by Sphingobacterium multivorum
1. Preparation of crude enzyme solution
Taking 5g of 1-2cm straw segments, adding 50mL of straw ligninase-producing culture medium, and sterilizing at 121 ℃ for 15 min. Inoculating Sphingobacterium multivorum into lignin enzyme production culture medium, culturing in30 deg.C incubator, and periodically supplementing water to maintain initial humidity. Adding 20ml of 0.2 mol.L into 1g of straw segment at the 6 th moment-1Acetic acid buffer solution (pH 5.0), placing on a constant temperature oscillator at 30 deg.C, and leaching for 2h to obtain leaching solution as crude enzyme solution.
2. Determination of Activity of manganese peroxidase, laccase and Lignin Catalase
To 3.4mL of 0.11 mol. L-1To a buffer solution of lactic acid-sodium lactate (pH 4.5) was added 0.1mL of 40 mmol/L-1MnSO of4And (3) preheating the solution in a constant-temperature water bath at 30 ℃ for 10min, adding 0.4mL of the crude enzyme solution prepared in the step (1), adding a hydrogen peroxide solution to start reaction, and placing the solution in an ice water bath to stop the reaction after 1 min. The absorbance change at a wavelength of 240nm was measured by an ultraviolet-visible spectrophotometer.
The fermentation method is the same as the fermentation method, the lignin enzyme production culture medium of the straws is fermented by the sphingosine bacillus polyphaga, and then a proper amount of straw sections are taken out and placed inIn the centrifugal tube, 20mL of 0.2 mol.L is added into each gram of straw section-1Acetic acid-sodium acetate buffer solution, placing in a constant temperature oscillator for leaching for 2h, and using the obtained leaching liquor as crude enzyme liquid for the following reaction process.
To 2.7mL of an acetic acid-sodium acetate buffer solution (pH 4.5), 0.2mL of 0.5 mmol/L was added-1The ABTS solution was preheated in a 30 ℃ constant temperature water bath for 10 minutes, and then 0.1mL of the crude enzyme solution was added thereto to start the reaction. After 1min, the reaction solution is put into an ice water bath to terminate the reaction, and an ultraviolet visible spectrophotometer is used for measuring the absorbance change value at the wavelength of 420 nm.
The detection result shows that the enzyme activities of laccase, manganese catalase and lignin catalase are respectively 0.57 U.mL in the fermentation liquor after 6 days of culture-1、0.47U·mL-1And 0.38 U.mL-1. Therefore, the sphingosine multivorum can produce a large amount of laccase, manganese catalase and lignin catalase, so that the lignin can be efficiently degraded.
Example 4 straw-degrading ability of cellulose-decomposing bacteria to Rice, wheat and corn
1. Preparation of sphingobacterium multivorum strain liquid
Sterilizing the liquid culture medium at 121 deg.C for 15min, and cooling to room temperature. Inoculating a bacterial liquid of sphingosine bacillus multivorus with 3 vol% into 200mL of the sterilized bacterial liquid culture medium, and culturing for 48h under the culture conditions of 150-.
2. Determination of straw degradation capability of sphingosine multivorum
The sphingosine bacillus multivorus seed liquid prepared in the preparation example is added into soil containing crop straws (straws of rice, wheat and corn respectively) according to the amount of 10mL of the bacterial seed liquid added into each kilogram of soil, the mixture is uniformly stirred, and the cultivation is carried out under the conditions that the environmental temperature is 20-30 ℃ and the water content is 15-30%, and the soil humidity is maintained. And after 21d, taking out the straw embedded in the straw, washing the straw with deionized water for 3 times, eluting soil and bacterial strains attached to the surface of the straw, putting the straw into an oven, drying the straw to constant weight, and weighing the mass of the straw after degradation. Finally, the weight loss ratio was calculated, where the sample weight loss ratio is (sample weight before degradation-sample weight after degradation)/sample weight before degradation × 100%. Comparing the degradation effect of the sphingosine bacillus multivorus on various straws, as shown in table 2, the data is the degradation effect data of the sphingosine bacillus multivorus on rice straws, wheat straws and corn straws.
TABLE 2
As shown in Table 2, the calculation results show that the degradation rates of the sphingosine dormitopsis on rice straw, wheat straw and corn straw are 73.26%, 60.99% and 67.13%, respectively, under the conditions of the present example.
The sphingosine bacillus multivorus provided by the embodiment of the invention has an obvious degradation effect on the straws in soil and water, can quickly decompose the straws and release nutrient substances, and solves a series of problems that the straw decomposition speed is low, the subsequent crop rooting is influenced and the like in the existing straw returning technology.
Although the invention has been described in detail with respect to the general description and the specific embodiments, it will be apparent to those skilled in the art that modifications and improvements may be made based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Sequence listing
<110> Anhui south Anhui tobacco, Inc., Beijing cereal and Runsheng science and technology, Inc
<120> sphingosine multivorum strain and application thereof
<130> GG19709890A
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 19
<212> DNA
<213> Artificial Sequence
<400> 1
agagtttgat cctggctca 19
<210> 2
<211> 19
<212> DNA
<213> Artificial Sequence
<400> 2
ggttaccttg ttacgactt 19
<210> 3
<211> 1421
<212> DNA
<213> Sphingobacterium multivorum
<400> 3
catgctttag gtacccccaa ctttcatggc ttgacgggcg gtgtgtacaa ggcccgggaa 60
cgtattcacc gcgtcattgc tgatacgcga ttactagcga atccaacttc atggggtcga 120
gttgcagacc ccaatccgaa ctgtgaatgg cttttagaga ttagcatcat attgctatgt 180
agctgcccgc tgtaccatcc attgtagcac gtgtgtagcc ccggacgtaa gggccatgat 240
gacttgacgt cgtccccacc ttcctctctg tttgcacagg cagtctgttt agagtcccca 300
ccactacatg ctggcaacta aacatagggg ttgcgctcgt tgcgggactt aacccaacac 360
ctcacggcac gagctgacga cagccatgca gcacctagtt tcgtgtcccg aaggacgagt 420
gcgtctctgc actcttcact aactttcaag cccgggtaag gttcctcgcg tatcatcgaa 480
ttaaaccaca tgctcctccg cttgtgcggg cccccgtcaa ttcctttgag tttcaccctt 540
gcgggcgtac tccccaggtg gataacttaa cgctttcgct tggacgctgg ctgtctatcg 600
ccaacatcga gttatcatcg tttagggcgt ggactaccag ggtatctaat cctgttcgat 660
ccccacgctt tcgtgcatca gcgtcaatac cagcttagtg agctgccttc gcaatcggag 720
ttctaagaca tatctatgca tttcaccgct acttgtctta ttccgcccac ttcaaatgga 780
ttcaagccca tcagtatcaa aggcactgcg atggttgagc caccgtattt cacccctgac 840
ttaataggcc gcctacgcac cctttaaacc caataaatcc ggataacgct cggatcctcc 900
gtattaccgc ggctgctggc acggagttag ccgatcctta ttcttccagt acattcagct 960
agatacacgt atctaggttt attcctggac aaaagcagtt tacaacccat agggcagtca 1020
tcctgcacgc ggcatggctg gttcaggctt ccgcccattg accaatattc cttactgctg 1080
cctcccgtag gagtctggtc cgtgtctcag taccagtgtg ggggattctc ctctcagagc 1140
ccctagacat cgtcgccttg gtaagccgtt accctaccaa ctagctaatg tcacgcgagc 1200
ccatctctat cctataaata tttaatcaac tgaacatgcg aactgttgat gttatgcggt 1260
gttaatctct ctttcgagag gctatccccc tgatagaggt aggttgctca cgcgttacgc 1320
acccgtgcgc cactctcacc atcttcgagc aagctctccg atggatcccg tccgacttgc 1380
atgtattagg cctgccgcta gcgttcatcc tgagccatga t 1421
Claims (7)
1. A cellulolytic bacterium, wherein said cellulolytic bacterium is a sphingobacterium (Sphingobacterium multivorum)Sphingobacterium multivorum) LWC-3, which is preserved in China general microbiological culture Collection center (CGMCC) within 22 days 5.2015, wherein the preservation address is No. 3 of Xilu-Beijing institute of microbiology, national academy of sciences, and the preservation number is CGMCC No. 10851.
2. A cellulose-degrading enzyme preparation comprising a fermentation broth of the bacterium sphingomyelinum according to claim 1.
3. The cellulose degrading enzyme preparation according to claim 2 comprising cellulase and/or hemicellulase and/or laccase and/or manganese catalase and/or lignin catalase.
4. The cellulose-degrading enzyme preparation according to claim 2 wherein the degradation substrate comprises sodium carboxymethyl cellulose and/or lignin.
5. Sphingobacterium multivorum (b) according to claim 1Sphingobacterium multivorum) The LWC-3 is applied to the degradation of cellulose.
6. Use according to claim 5, wherein the cellulose is straw.
7. The use of claim 6, wherein the straw is at least one of rice straw, wheat straw and corn straw.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911378511.6A CN111172062B (en) | 2019-12-27 | 2019-12-27 | Sphingobacterium multivorum and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911378511.6A CN111172062B (en) | 2019-12-27 | 2019-12-27 | Sphingobacterium multivorum and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111172062A CN111172062A (en) | 2020-05-19 |
| CN111172062B true CN111172062B (en) | 2022-06-03 |
Family
ID=70618914
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911378511.6A Active CN111172062B (en) | 2019-12-27 | 2019-12-27 | Sphingobacterium multivorum and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111172062B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115820490B (en) * | 2022-11-29 | 2023-06-27 | 江西省科学院生物资源研究所 | Sphingobacterium polycephalum, microbial inoculum and application thereof in preventing and treating kiwi soft rot |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001178447A (en) * | 1999-12-24 | 2001-07-03 | Shinohara Seiki Kk | Microbial group having agrochemical-decomposing ability, compost containing the microbial group and method and apparatus for producing the compost |
| CN101885624A (en) * | 2010-07-16 | 2010-11-17 | 湖北金帝农业科技有限公司 | Decomposition maturing agent for degrading straw |
| RU2010154237A (en) * | 2010-12-29 | 2012-07-10 | Государственное научное учреждение Татарский научно-исследовательский институт агрохимии и почвоведения Российской академии сельско | STRAIN SPHINGOBACTERIUM MULTIVORUM USED FOR OBTAINING BACTERIAL FERTILIZER FOR TOMATOES AND CUCUMBERS |
| WO2013188844A2 (en) * | 2012-06-15 | 2013-12-19 | Microvi Biotech Inc. | Bioconversion processes and apparatus |
-
2019
- 2019-12-27 CN CN201911378511.6A patent/CN111172062B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001178447A (en) * | 1999-12-24 | 2001-07-03 | Shinohara Seiki Kk | Microbial group having agrochemical-decomposing ability, compost containing the microbial group and method and apparatus for producing the compost |
| CN101885624A (en) * | 2010-07-16 | 2010-11-17 | 湖北金帝农业科技有限公司 | Decomposition maturing agent for degrading straw |
| RU2010154237A (en) * | 2010-12-29 | 2012-07-10 | Государственное научное учреждение Татарский научно-исследовательский институт агрохимии и почвоведения Российской академии сельско | STRAIN SPHINGOBACTERIUM MULTIVORUM USED FOR OBTAINING BACTERIAL FERTILIZER FOR TOMATOES AND CUCUMBERS |
| WO2013188844A2 (en) * | 2012-06-15 | 2013-12-19 | Microvi Biotech Inc. | Bioconversion processes and apparatus |
Non-Patent Citations (3)
| Title |
|---|
| Diego Javier Jimenez et al.."Defining the eco-enzymological role of the fungal strainConiochaetasp. 2T2.1 in a tripartite lignocellulolytic microbial consortium".《FEMS Microbiology Ecology》.2019,第96卷(第1期),第1-12页. * |
| Larisa Cortes-Tolalpa et al.."Bacterial Synergism in Lignocellulose Biomass Degradation-Complementary Roles of Degraders As Influenced by Complexity of the Carbon Source".《Frontiers in Microbiology》.2017,第8卷第1-14页. * |
| 刘淑霞 等."几种纤维素分解菌在有机质转化中的作用".《农业环境科学学报》.2008,第27卷(第3期),991- 996. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111172062A (en) | 2020-05-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106399209B (en) | Composite microbial inoculum for degrading high-grease kitchen waste and preparation method thereof | |
| US20230242868A1 (en) | Pseudomonas graminis strain capable of degrading cellulose at low temperature and use thereof | |
| CN111073839B (en) | Siam bacillus, microbial inoculum and application thereof | |
| CN110042072A (en) | A kind of aflatoxin degradation B1Bacterial strain and its application | |
| CN114480205B (en) | Bacillus amyloliquefaciens and application thereof in brewing of solid-state fermentation vinegar | |
| CN110484462B (en) | Novel species of genus Shen-shi and application thereof | |
| CN115851545A (en) | Korean bacillus and culture medium for improving activity of enzyme produced by same | |
| CN111172062B (en) | Sphingobacterium multivorum and application thereof | |
| CN111154661B (en) | Complex microbial inoculant and application thereof | |
| CN112592862A (en) | Preparation method and application of aerobic fermentation salt-tolerant composite microbial agent | |
| CN104560817A (en) | Thermophilic bacillus licheniformis UTM102 for producing phytase and application of thermophilic bacillus licheniformis UTM102 | |
| CN102173879B (en) | Method for producing biological potassium fertilizer by utilizing cellulose fermented waste mycelium and biogas residue | |
| CN104560815B (en) | Bacillus licheniformis with azo compound degradation activity and application thereof | |
| CN103255062B (en) | Aspergillus niger XL-1, and microorganism preparation thereof, and application of aspergillus niger XL-1 in straw degradation | |
| CN111733087B (en) | Lacquercus humilis for producing laccase and application thereof | |
| CN114717125B (en) | Thermophilic bacillus licheniformis AMCC101380 and application thereof in high-temperature composting of tail vegetables | |
| CN116875501A (en) | Kitchen waste decomposing degradation microbial inoculum and preparation method and application thereof | |
| CN110452852B (en) | Farmer market biomass waste fermentation inoculant and preparation method thereof | |
| CN106348884A (en) | Production method of bacillus megaterium and humic acid containing liquid water-soluble fertilizer | |
| CN112625960A (en) | Agrobacterium DP3, microbial inoculum thereof and application thereof in field of biological fertilizer preparation | |
| CN110317755B (en) | Low-temperature-resistant cellulolytic pseudomonas menbergii and application thereof | |
| CN113717882B (en) | Geobacillus galactose BWTGW1.1 and application thereof | |
| CN110527643A (en) | One plant of soil Psychrobacter and its application in agricultural solid waste low temperature aerobic fermentation | |
| CN114317330B (en) | Screening and application of cellulose degrading bacteria | |
| CN116515795B (en) | Application of Aspergillus tubingensis in preparing phytase and/or degrading phytic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 242000 No.72, Aofeng Middle Road, Xuanzhou District, Xuancheng City, Anhui Province Applicant after: ANHUI WANNAN TOBACCO LEAF Co.,Ltd. Applicant after: Beijing Tianhe Ruizhi Technology Co.,Ltd. Address before: 242000 No.72, Aofeng Middle Road, Xuanzhou District, Xuancheng City, Anhui Province Applicant before: ANHUI WANNAN TOBACCO LEAF Co.,Ltd. Applicant before: BEIJING HEHERUNSHENG TECHNOLOGY Co.,Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |